中国仓鼠V79细胞系的oua~R和oua~S细胞蛋白质组分的双向电泳比较

作者用聚丙烯酰胺凝胶双向电泳,对中国仓鼠V79细胞系的乌本苷抗性突变型(oua~R)和野生型(oua~S)细胞的蛋白质组分进行比较,选择20个高度重复、分布均匀的肽斑作为参考标记,发现一些多肽在其位置、大小及密度上表现出差异,值得注意的是4小区(pI/MW为5.0/28～30kd)和6小区(pI/MW为6.3/39kd)的变化明显,提示这些多肽可能对V79细胞的oua~R是特异的,也许可作为oua~R细胞的特征性遗传标记。

COMPARATIVE STUDY ON PROTEIN COMPONENTS OF OUABAIN RESISTANT MUTANT AND WILD CELLS OF CHINESE HAMSTER V79 CELL LINE BY TWO-DIMENSIONAL GEL ELECTROPHORESIS

In the sense of gene expression, theprimary and direct change in mutant cellsshould be involved in the protein compo-nent. Two-dimensional polyacrylamide gelelectrophoresis has been used to compare theprotein components of ouabain resistantmutant cells (oua~R) and sensitive wild typecells(oua~s)of chinese hamster lung cell lineV_(79). The procedure is based on O'Farrell'smethod and we have made some modifica-tions. In order to make the study reliable,we took twenty polypeptide spots as a re-ference marker. These marker spots couldalways be reproduced in a series of parallelexperiments with both mutant and wildcells. In addition, these markers covereddifferent small areas, representing dif-ferent isoelectric point (pI) and molecularweight (MW) on the electrophoretogram. With reference to these marker spots.some polypeptide spots showed varia-tions in their position, size and density.It is notable that the changes appeared inthe small area 4 (pI/MW:5.0/28-30kd) andthe small area (pI/MW:6.3/39kd) is highlyreproducible and significant, so that thesechanges might be specific to ouabain resis-tant mutant of V_(79) cells. Since the level of genetic polymorphismof mammalian cells on two-dimensional gelelectrophoresis is quite low, these specificchanges might reflect the differences ingene structure and expression. This primary study should enhance theusefulness of two-dimensional gel electro-phoresis for mutagenesis research with cul-tured mammlian somatic cells.